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Annals of Hematology

, Volume 98, Issue 5, pp 1083–1093 | Cite as

Features, reason for testing, and changes with time of 583 paroxysmal nocturnal hemoglobinuria clones from 529 patients: a multicenter Italian study

  • Elisa Cannizzo
  • Maddalena Raia
  • Maria Stefania De Propris
  • Anna Triolo
  • Barbara Scarpati
  • Anna Marfia
  • Alessandra Stacchini
  • Francesco Buccisano
  • Francesco Lanza
  • Antonio Regazzoli
  • Angela Michelutti
  • Simone Cesaro
  • Cinzia Armentano Conte
  • Laura Vanelli
  • Elisabetta Tedone
  • Paola Omedè
  • Maria Matilde Ciriello
  • Roberto Caporale
  • Virginia Catinella
  • Giorgia Pantano
  • Clorinda De Rosa
  • Catia Lo Pardo
  • Giovanni Poletti
  • Francesca Ulbar
  • Maria Cristina Pavanelli
  • Laura Del Pup
  • Virginia Ottaviano
  • Anna Maria Santonocito
  • Chiara Bartocci
  • Elisa Boscaro
  • Marcella Arras
  • Rachele Amodeo
  • Anna Mestice
  • Bianca Oliva
  • Luisa Ferrari
  • Teodora Statuto
  • Fiorella D’Auria
  • Graziano Pianezze
  • Donatella Tanca
  • Feliciano Visconte
  • Fabiana Rubba
  • Pellegrino Musto
  • Massimo Geuna
  • Arianna Gatti
  • Bruno BrandoEmail author
  • Luigi Del Vecchio
Original Article

Abstract

In this study, we aimed at disclosing the main features of paroxysmal nocturnal hemoglobinuria (PNH) clones, their association with presentation syndromes, and their changes during follow-up. A large-scale, cooperative collection (583 clones from 529 patients) of flow cytometric and clinical data was entered into a national repository. Reason for testing guidelines were provided to the 41 participating laboratories, which followed the 2010 technical recommendations for PNH testing by Borowitz. Subsequently, the 30 second-level laboratories adopted the 2012 guidelines for high-resolution PNH testing, both upon order by the local clinicians and as an independent laboratory initiative in selected cases. Type3 and Type2 PNH clones (total and partial absence of glycosyl-phosphatidyl-inositol-anchor, respectively) were simultaneously present in 54 patients. In these patients, Type3 component was sevenfold larger than Type2 (p < 0.001). Frequency distribution analysis of solitary Type3 clone size (N = 442) evidenced two discrete patterns: small (20% of peripheral neutrophils) and large (> 70%) clones. The first pattern was significantly associated with bone marrow failure and myelodysplastic syndromes, the second one with hemolysis, hemoglobinuria, and thrombosis. Pediatric patients (N = 34) showed significant preponderance of small clones and bone marrow failure. The majority of PNH clones involved neutrophils, monocytes, and erythrocytes. Nevertheless, we found clones made exclusively by white cells (N = 13) or erythrocytes (N = 3). Rare cases showed clonal white cells restricted only to monocytes (6 cases) or neutrophils (3 cases). Retesting over 1-year follow-up in 151 cases showed a marked clone size increase in 4 cases and a decrease in 13, demonstrating that early breaking-down of PNH clones is not a rare event (8.6% of cases). This collaborative nationwide study demonstrates a clear-cut difference in size between Type2 and Type3 clones, emphasizes the existence of just two classes of PNH presentations based on Type3 clone size, depicts an asymmetric cellular composition of PNH clones, and documents the possible occurrence of changes in clone size during the follow-up.

Keywords

Paroxysmal nocturnal hemoglobinuria Flow cytometry Myelodysplastic syndromes Hemolytic anemia Atypical thrombosis Aplastic anemia 

Abbreviations

AA

Aplastic anemia

BMF`

Bone marrow failure

CEINGE

Institute for Advanced Biotechnologies, Naples University

DAT

Direct anti-globulin test

FCM

Flow cytometry

FLAER

Fluorescent aerolysin

GPI

Glycosyl-phosphatidyl-inositol

HA

Hypoplastic anemia

ICD

International classification of the diseases

MDS

Myelodysplastic syndromes

PNH

Paroxysmal nocturnal hemoglobinuria

RFT

Reason for testing

SCAPE

Standardization of a cytometric approach to PNH clone evaluation

Notes

Compliance with ethical standards

All studies in this paper were performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments. All procedures followed were in accordance with the Helsinki Declaration of 1975, as revised in 2008. Although this type of study did not require a formal clearance by ethical committees, all persons gave their informed consent prior to the inclusion of their data in the national repository.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

277_2019_3644_MOESM1_ESM.docx (85 kb)
ESM 1 (DOCX 84 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Elisa Cannizzo
    • 1
  • Maddalena Raia
    • 2
    • 3
  • Maria Stefania De Propris
    • 4
  • Anna Triolo
    • 5
  • Barbara Scarpati
    • 6
  • Anna Marfia
    • 7
  • Alessandra Stacchini
    • 8
  • Francesco Buccisano
    • 9
  • Francesco Lanza
    • 10
  • Antonio Regazzoli
    • 11
  • Angela Michelutti
    • 12
  • Simone Cesaro
    • 13
  • Cinzia Armentano Conte
    • 14
  • Laura Vanelli
    • 15
  • Elisabetta Tedone
    • 16
  • Paola Omedè
    • 17
  • Maria Matilde Ciriello
    • 18
  • Roberto Caporale
    • 19
  • Virginia Catinella
    • 20
  • Giorgia Pantano
    • 21
  • Clorinda De Rosa
    • 22
  • Catia Lo Pardo
    • 23
  • Giovanni Poletti
    • 24
  • Francesca Ulbar
    • 25
  • Maria Cristina Pavanelli
    • 26
  • Laura Del Pup
    • 27
  • Virginia Ottaviano
    • 28
  • Anna Maria Santonocito
    • 29
  • Chiara Bartocci
    • 30
  • Elisa Boscaro
    • 31
  • Marcella Arras
    • 32
  • Rachele Amodeo
    • 33
  • Anna Mestice
    • 34
  • Bianca Oliva
    • 35
  • Luisa Ferrari
    • 36
  • Teodora Statuto
    • 37
  • Fiorella D’Auria
    • 37
  • Graziano Pianezze
    • 38
  • Donatella Tanca
    • 39
  • Feliciano Visconte
    • 2
    • 3
  • Fabiana Rubba
    • 40
  • Pellegrino Musto
    • 37
  • Massimo Geuna
    • 41
  • Arianna Gatti
    • 42
  • Bruno Brando
    • 42
    Email author
  • Luigi Del Vecchio
    • 2
    • 3
    • 43
  1. 1.Servizio Medicina Trasfusionale ed EmatologiaRagusaItaly
  2. 2.CEINGE Biotecnologie AvanzateNaplesItaly
  3. 3.Dipartimento di Medicina Molecolare e Biotecnologie MedicheUniversità Federico IINaplesItaly
  4. 4.Divisione di Ematologia, Dipartimento di Biotecnologie Cellulari ed EmatologiaUniversità La SapienzaRomeItaly
  5. 5.Ematologia e Trapianto di Midollo Osseo, P.O. FerrarottoAOU Vittorio EmanueleCataniaItaly
  6. 6.Servizio di Immunologia e Medicina TrasfusionaleASST Grande Ospedale Metropolitano NiguardaMilanItaly
  7. 7.Divisione di EmatologiaOspedali Riuniti Villa Sofia-CervelloPalermoItaly
  8. 8.UOC Anatomia Patologica Generale e Oncogenetica Molecolare, PO MolinetteAOU Città della Salute e della ScienzaTorinoItaly
  9. 9.UOC EmatologiaUniversità Tor VergataRomeItaly
  10. 10.UO di EmatologiaIstituti OspitalieriCremonaItaly
  11. 11.Laboratorio AnalisiAO Spedali CiviliBresciaItaly
  12. 12.Divisione di Ematologia e Trapianto di MidolloAOU UdineUdineItaly
  13. 13.Oncoematologia PediatricaAOU VeronaVeronaItaly
  14. 14.Divisione di Ematologia e Trapianto Cellule StaminaliAO San CamilloRomeItaly
  15. 15.Dip OncoematologicoIRCCS S. MatteoPaviaItaly
  16. 16.UOC Ematologia2AOU S. MartinoGenoaItaly
  17. 17.Lab Citometria, UOC EmatologiaAOU Città della Salute e della ScienzaTorinoItaly
  18. 18.Laboratorio AnalisiAO SS. Antonio e BiagioAlessandriaItaly
  19. 19.Area Diagnostica di Laboratorio, Dip ServiziAOU CareggiFlorenceItaly
  20. 20.UOC Immunoematologia e Centro TrasfusionaleAO S SpiritoPescaraItaly
  21. 21.Dipartimento Interaziendale Medicina di LaboratorioPoliclinico UniversitarioPadovaItaly
  22. 22.Lab Citometria, UOC MedicinaEnte Ecclesiastico Ospedale F. MiulliAcquaviva delle FontiItaly
  23. 23.Centro TrasfusionaleAORN CardarelliNaplesItaly
  24. 24.Laboratorio EmatologiaLaboratorio AVRPievesestinaItaly
  25. 25.Laboratorio di Immunologia, Istituto di Ematologia L. & A. SeràgnoliAOU S.Orsola-MalpighiBolognaItaly
  26. 26.SS Citometria a flusso, UOC Anatomia Patologica, PO S. AndreaVercelliItaly
  27. 27.Laboratorio di Ematologia, Servizio ImmunotrasfusionaleTrevisoItaly
  28. 28.Laboratorio di CitofluorimetriaUO di Ematologia Universitaria AOUPPisaItaly
  29. 29.UOC Ematologia con Laboratorio di Biologia Molecolare, PO San VincenzoTaorminaItaly
  30. 30.Dipartimento Ospedaliero di Medicina InternaAOU Ospedali RiunitiAnconaItaly
  31. 31.Dipartimento di MedicinaEmatologia e Immunologia Clinica Policlinico UniversitarioPadovaItaly
  32. 32.CTMO, PO Binaghi, ASLCagliariItaly
  33. 33.Sezione di Citometria, UOC Diagnostica di laboratorio, AO S. AndreaUniversità La SapienzaRomeItaly
  34. 34.Dipartimento Ematologia con TrapiantoAOU PoliclinicoBariItaly
  35. 35.UOC Patologia Clinica, AO Bianchi Melacrino MorelliReggio CalabriaItaly
  36. 36.Lab Ematologia, AOUFerraraItaly
  37. 37.Direzione Scientifica, IRCCS CROBRionero in VultureItaly
  38. 38.Ematologia, Ospedale S MartinoBellunoItaly
  39. 39.Settore Ematologia, Citometria e Coagulazione, Ospedale LavagnaLavagnaItaly
  40. 40.DAI Sanità PubblicaAOU Federico IINaplesItaly
  41. 41.Laboratorio di Patologia Oncoematologica, Anatomia PatologicaAO Ordine MaurizianoTorinoItaly
  42. 42.Hematology Laboratory and Transfusion Center, Western Milan Area Hospital ConsortiumLegnano General HospitalLegnanoItaly
  43. 43.DAI Medicina TrasfusionaleAOU Federico IINaplesItaly

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